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Nanosized polymer films can be constructed via alternative electrostatic adsorption of polyelectrolytes. The adsorption is to a great extent determined by the state and structure of initial surface and/or the pre-adsorbed layers. The same rule is apparently valid for electrostatic adsorption of biomolecules used for their immobilization. This is specifically important for a design and fabrication of bioanalytical devices, e.g., biosensors, where the amount of biomolecules incorporated and the strength of their binding influence on basic characteristics, such as activity and operation stability, of these bioanalytical surfaces. To demonstrate this, different fabrication regimes and properties of polymer/biomolecule thin films adsorbed onto conductive substrates were examined. The films were formed via two-steps, sequential adsorption of a polymeric component followed by the biomolecule adsorption under different pH/salt composition and temperature regimes. Strong/weak linear polyelectrolytes, amphiphilic ionogenic diblock copolymers or microgels were considered as polymeric components to be interacted with biomolecules, like enzymes or hemeproteins. Monoenzymatic systems with uniformly distributed biomaterial or bienzymatic systems with spatially separated enzymes are discussed. Specific aplications to the field of biosensors are discussed. (O58, p. 110)